Preparation method of cell suspension with melanocyte activity
A technology of melanocytes and cell suspensions, applied in the field of preparation of cell suspensions, can solve the problems of cell inactivation, large differences in skin color, weak proliferation ability, etc., achieve high activity, promote adhesion, and repair wounds
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[0016] see figure 1 , a preferred embodiment of the present invention provides a method for preparing a cell suspension with melanocyte activity, comprising the following steps:
[0017] Step S11, taking a blade-thick skin slice (ie, superficial skin slice) of healthy skin.
[0018] Specifically, perform skin disinfection on the patient's skin-taking site, and use a skin-taking machine to take blade-thick skin slices of the patient's healthy skin. Wherein, the thickness of the blade thickness sheet is 200-500 μm.
[0019] Step S12, placing the blade-thick skin piece in a digestive solution for digestion.
[0020] Specifically, soak the blade-thick skin piece in the digestive solution for 15-45min to digest, wherein, the amount of the digestive juice per square centimeter of the blade-thick skin piece is 3-7mL, the digestion temperature is 37°C, and the digestion time is 37°C. Use a scraper to scrape the epidermis in the described blade-thickness skin sheet after using scrap...
Embodiment 1
[0036] The first step is to use a skin remover to take a piece of healthy skin with a blade thickness. Wherein, the thickness of the blade-thickness skin is 200 μm, and the size of the blade-thickness skin is 1×1 cm.
[0037] In the second step, soak the blade-thickness skin in 5mL of digestive juice and digest at 37°C for 15-45min. Scrape off the epidermis, then proceed to the next step; if the epidermis cannot be easily scraped off, then put it back into the digestive solution to continue digestion until the epidermis can be scraped off, wherein the total time of digestion cannot More than 45min. Wherein, the digestive juice includes trypsin with a concentration of 0.5% and EDTA with a concentration of 0.05%.
[0038] In the third step, after washing the digested epidermis three times in the first nutrient solution, place the epidermis upwards on the operating table, add a little second nutrient solution dropwise, and gently scrape the epidermis with the scraper , until t...
Embodiment 2
[0042] The difference between embodiment 2 and embodiment 1 is:
[0043] In the first step, the thickness of the blade-thickness skin is 300 μm, and the size of the blade-thickness skin is 2×1 cm.
[0044] In the second step, the blade-thick skin piece was soaked in 10 mL of digestive solution, which included trypsin at a concentration of 0.5%, EDTA at a concentration of 0.05%, and rh-EGF at a concentration of 8 ng / mL .
[0045]In the third step, the first nutrient solution is Ringer's buffer. The second nutrient solution includes mixed medium (obtained by mixing DMEM medium and Ham's F-12 medium at a volume ratio of 3:1), hydrocortisone with a concentration of 1 μM, insulin with a concentration of 0.1 μM, and a concentration of 10 μM L-carnitine, 10 μM L-serine, 15 μM linoleic acid, 1 μM isoproterenol, 2 ng / mL KGF and 4% fetal bovine serum.
[0046] In the fifth step, the cell concentration in the cell suspension with melanocyte activity is 1×10 7 / mL.
[0047] The cell ...
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